Journal: bioRxiv

Article Title: Perineuronal Net and Inhibitory Synapse Remodeling on Striatal Fast-spiking Interneurons by Chronic Alcohol Exposure

doi: 10.1101/2025.07.08.663744

Figure Lengend Snippet: A ) schematic and timeline of CIE vapor exposure paradigm. B ) Schematic of whole-cell patch clamp recording from an FSI. C ) CIE dramatically reduced inhibitory synaptic transmission onto FSIs (N=6 mice per group, 3M 3F). Inset: representative electrically evoked inhibitory postsynaptic currents (eIPSCs) from control (black) and CIE (blue) treated mice (scale bars 500pA, 100ms). D ) However, we observed no change in electrically-evoked excitatory postsynaptic currents (eEPSCs) onto FSIs (N=6 mice per group, 3M 3F). E, F ) Spontaneous inhibitory synaptic transmission onto dorsal striatum FSIs (N=10 mice per group, 5M 5F). Inset: representative traces from control (black) and CIE (blue) treated mice (scale bar 100pA, 2s). CIE reduced the frequency of spontaneous inhibitory events ( E, F ) but did not impact event amplitude ( G ). H ) CIE did not impact FSI dendrite branch complexity (N=10 cells, 5 mice per group, 3M 2F). Representative dye-filled FSIs from control (left) and CIE (right) treated mice (Sholl ring radius = 10µm increments). All data represented as mean + SEM. * P<0.01. Portions of this figure were created with BioRender.com.

Article Snippet: Dorsolateral striatum FSIs were fluorescently visualized using Eclipse FN1 Nikon microscope and Intenslight C-HGFIE (Nikon).

Techniques: Patch Clamp, Transmission Assay, Control

Journal: bioRxiv

Article Title: Perineuronal Net and Inhibitory Synapse Remodeling on Striatal Fast-spiking Interneurons by Chronic Alcohol Exposure

doi: 10.1101/2025.07.08.663744

Figure Lengend Snippet: A ) Schematic of injection of a virus expressing channelrhodopsin (ChR2) into the globus pallidus. B ) CIE reduced optically-evoked inhibitory currents from the GP onto FSIs but ( C ) had no impact on release probability (N=8 mice per group, 4M 4F) Inset: representative traces from control (black) and CIE (blue) treated mice (scale bar 500 pA, 100 ms). CIE reduced the ( D ) frequency but not the ( E ) amplitude of asynchronous inhibitory current events from the GP. Inset: representative traces from control (black) and CIE (blue) treated mice (scale bar 200 pA, 10 ms). F ) Schematic of injection of virus expressing ChR2 into the RTN. G ) CIE reduced optically-evoked inhibitory currents from the RTN but ( H ) had no impact on release probability (N=8 mice per group, 4M 4F; scale bar 500pa, 100ms). I ) CIE reduced the frequency of asynchronous inhibitory current events but ( J ) did not impact asynchronous current event amplitude (N= 6 mice per group, 3M 3F; scale bar 200 pA, 20 ms). K ) Schematic of injection of cre-dependent virus expressing tetanus light chain toxin (TeLC) into the GP and RTN and retrograde AAV expressing cre-recombinase in the dorsal striatum. L ) Experimental timeline of Drinking in the Dark (DID) paradigm with schematic of mouse drinking ethanol (20%) or sucrose (2%). M ) TelC silencing of inhibitory transmission from the GP and RTN did not impact sucrose consumption. N ) However, TelC silencing of inhibitory transmission increased voluntary ethanol consumption. All data represented as mean + SEM. *P < 0.05, **P < .01. Portions of this figure were created with BioRender.com.

Article Snippet: Dorsolateral striatum FSIs were fluorescently visualized using Eclipse FN1 Nikon microscope and Intenslight C-HGFIE (Nikon).

Techniques: Injection, Virus, Expressing, Control, Transmission Assay

Journal: bioRxiv

Article Title: Perineuronal Net and Inhibitory Synapse Remodeling on Striatal Fast-spiking Interneurons by Chronic Alcohol Exposure

doi: 10.1101/2025.07.08.663744

Figure Lengend Snippet: A ) Schematic of viral injection of virus expressing a cre-dependent Gephyrin-GFP intrabody into parvalbumin (PV)-cre mice. B ) Representative image of an FSI expressing gephyrin-GFP intrabody (green) colocalized to an immunofluorescent marker for vesicular GABA transporter (VGAT) puncta. VGAT positive puncta were masked onto the GFP signal and colocalized signals (white dots) were labeled with a dot function model to quantify GABAergic synapses (scale bar 10µm). (right) Representative fluorescent image of proximal neuronal branch from control and CIE treated mice depicting colocalized presynaptic VGAT (purple) puncta onto gephyrin (green)-positive postsynaptic elements (scale bar 1µm). CIE dramatically reduced the density of colocalized VGAT puncta onto somata ( C ), proximal dendrites ( D ), and distal dendrites ( E ) (N=8 mice per group, 4M 4F). F ) Schematic of whole-cell patch clamp recording from an FSI during photo-uncaging of RuBi-GABA. G ) Representative traces of photo-uncaged GABA currents from FSIs of control (left) and CIE (right) treated mice at increasing stimulus intensities (scale bar = 200pA, 100ms). H ) CIE did not impact photo-uncaged IPSCs compared to control treated mice (n=8 mice per group, 4M 4F). All data represented as mean + SEM. *P < .05, **p<.01. Portions of this figure were created with BioRender.com

Article Snippet: Dorsolateral striatum FSIs were fluorescently visualized using Eclipse FN1 Nikon microscope and Intenslight C-HGFIE (Nikon).

Techniques: Injection, Virus, Expressing, Marker, Labeling, Control, Patch Clamp

Journal: bioRxiv

Article Title: Perineuronal Net and Inhibitory Synapse Remodeling on Striatal Fast-spiking Interneurons by Chronic Alcohol Exposure

doi: 10.1101/2025.07.08.663744

Figure Lengend Snippet: A ) Immunofluorescent image of parvalbumin+ FSIs (red) and WFA+ perineuronal nets (green) depicting net enriched (PNN+) and net poor (PNN-) FSIs (scale bar 100µm). B,C ) Fluorescent intensity profile of PNN+ and PNN-FSIs. D) CIE reduced the percentage of net enriched FSIs but ( E ) does not reduce the number PV+ FSIs (N=10 mice per group, 5M 5F). CIE reduced the expression of PNN proteins ( F ) aggrecan and ( G ) HAPLN1 in the dorsolateral striatum (N=8 mice per group, 4M 4F). H ) Representative confocal fluorescent images of PV+ FSI (black) and WFA+ PNN (green) from control (top) and CIE (bottom) treated mice (scale bar 15µm). I ) Top: 3D volume reconstruction and filament model of WFA fluorescent signal from control (top) and CIE (bottom) treated mice. J ) CIE reduced the volume of WFA signal (top) surrounding FSI somata and reduced the length of WFA signal (bottom) surrounding FSI proximal branches (N=10 mice per group, 5M 5F). All data represented as mean + SEM. *P < 0.05. ** P < 0.01.

Article Snippet: Dorsolateral striatum FSIs were fluorescently visualized using Eclipse FN1 Nikon microscope and Intenslight C-HGFIE (Nikon).

Techniques: Expressing, Control

Journal: bioRxiv

Article Title: Perineuronal Net and Inhibitory Synapse Remodeling on Striatal Fast-spiking Interneurons by Chronic Alcohol Exposure

doi: 10.1101/2025.07.08.663744

Figure Lengend Snippet: A ) schematic of whole-cell patch recording from PNN enriched (PNN+) or PNN poor (PNN-) FSI identified by WFA stain after recordings. B ) PNN+ FSIs exhibited larger eIPSC amplitudes compared to undetected PNN-FSIs (N=11 PNN+ cells, 5M 6F; 6 PNN-cells, 3M 3F). Inset: representative traces from PNN+ (pink) and PNN+ (black) FSIs (scale bar 1nA, 100ms). C ) The presence of PNNs did not impact GABA release probability onto FSIs. D-F ) Spontaneous IPSC events (sIPCSs) recorded from FSIs (N=11 PNN+ cells, 5M 6F; 6 PNN-cells, 3M 3F). Inset: representative traces from PNN+(pink) and PNN-(black) FSIs (scale bar 100pA, 1s). D,E ) PNN+ FSIs exhibited a greater frequency of spontaneous IPSC events but ( F ) no change in IPSC amplitude compared to PNN-FSIs. G ) Schematic of unilateral microinjection of chondroitinase ABC (ChABC) into the dorsolateral striatum to enzymatically degrade PNNs. H ) Degrading PNNs with ChABC reduced eIPSC event amplitude onto FSIs but ( I ) did not impact GABA release probability (N=10 mice, 5M 5F; scale bar 1nA, 100ms). J,K ) Degrading PNNs with ChABC reduced the frequency of sIPSC events onto FSIs but ( L ) did not impact event amplitude (N=8 mice, 4M 4F; scale bar 100pA, 1s). M ) Schematic of viral injection of cre-dependent Gephyrin-GFP intrabody into PV-cre mice followed by ChABC. N) Degrading PNNs with ChABC did not reduce inhibitory synapses onto somata but ( O ) did reduce inhibitory synapse number on proximal dendrites (N= 13 mice, 6M 7F). P ) Schematic of whole-cell patch clamp recording during photo-uncaging of RuBi-GABA onto FSIs treated with ChABC. Q ) Representative traces of photo-uncaged GABA postsynaptic currents on control (left) and ChABC (right) treated FSIs at increasing stimulus intensity (scale bar = 200pA, 100ms). G ) There was no change in photo-uncaged IPSC event amplitude between control (black) and ChABC (pink) FSIs (N=8 mice per group, 4M 4F). All data represented as mean + SEM. *P < 0.05. ** P < 0.01. Portions of this figure were created with BioRender.com

Article Snippet: Dorsolateral striatum FSIs were fluorescently visualized using Eclipse FN1 Nikon microscope and Intenslight C-HGFIE (Nikon).

Techniques: Staining, Microinjection, Injection, Patch Clamp, Control